压力对猪眼小梁细胞表达ELAM-1、MMPs/TIMPs的影响

Effect of pressure on expressions of ELAM-1,MMP-2,MMP-3 and TIMP-2 in porcine trabecular meshwork cells cultured in vitro

目的培养猪眼小梁细胞,研究压力对体外培养的猪眼小梁内皮细胞白细胞黏附分子-1(ELAM-1)、基质金属蛋白酶MM P-2、MM P-3及基质金属蛋白酶组织抑制剂TIM P-2表达的影响。方法培养猪眼小梁细胞并鉴定,建立细胞水平的青光眼模型,对传第3代猪眼小梁细胞分别施加20、40、60、80mmHg压力作为实验组,0mmHg为对照组。培养6h后行ELAM-1免疫组织化学SP法染色,培养24h后行MM P-2、MM P-3和TIM P-2免疫组织化学SP法染色,并对染色结果进行统计学分析。结果培养细胞为猪眼小梁细胞,正常小梁细胞不表达ELAM-1,压力为40、60、80mmHg时,ELAM-1的表达与0、20mmHg组相比表达明显增加。正常小梁细胞可以少量表达MM P-2、MM P-3及TIM P-2。压力为40、60mmHg时,MM P-2、TIM P-2的表达与0、20、80mmHg相比表达明显增加,压力不影响小梁细胞MM P-3的表达。结论一定范围内压力的变化可以促进猪眼小梁细胞表达ELAM-1,可以改变MM Ps/TIM Ps之间的平衡状态,进而影响小梁细胞外基质(ECM)的代谢,改变房水外流阻力,ELAM-1、MM Ps在青光眼的发病中可能发挥重要作用。

Objective To investigate the effect of pressure on expressions of endothelial leukocyte adhesion molecule-1 （ELAM-1）, matrix metalloproteinase-2 （ MMP-2 ）, MMP-3 and Tissue Inhibitor of MetaUoproteinase-2 （ TIMP-2 ） in porcine trabecular cells cultured in vitro. Methods Porcine trabecular cells were cultured, passaged and identified. The third passage generation trabecular cells were treated under pressure of 20, 40, 60 and 80 mmHg（ 1 kPa =7.5 ram- Hg） in the treatment group to establish a cell-level glaucoma model, while no pressure（0 mmHg） was given to the con- trol group. Immunocytochemical staining was used to observe expression of ELAM-1 after 6 hours＇ culture, and expres- sions of MMP-2, MMP-3 and TIMP-2 after 24 hours＇ culture. Staining results were statistically analyzed. Results Cultured cells were identified as porcine trabecular cells. Expression of ELAM-1 was not detected in trabecular cells of the control group, while expressions of MMP-2, MMP-3 and TIMP-2 were observed in small quantity. When pressure was 40, 60 or 80 mmHg, expression of ELAM-1 significantly increased compared with the 0 mmHg group and 20 ram- Hg group. When pressure was 40 or 60 mmHg, expressions of MMP-2 and TIMP-2 significantly increased compared with the 0 mmHg group, 20 mmHg group and 80 mmHg group. Expression of MMP-3 had no significant variation. Conclusion A rise of pressure can induce an increase in expressions of ELAM-1, MMP-2 and TIMP-1 in cultured por- cine trabecular cells, causes unbalance of MPs/TIMPs, affects metabolism of extracellularmatrix（ECM） and increasesaqueous outflow. Expressions of EL AM-1 and MMPs may contribute to the pathogenesis of glaucoma.